移植分泌神经营养因子-3的人胚神经干细胞对大鼠脊髓损伤后功能恢复的影响

目的探索Lentivirus介导神经营养因子(NT-3)转染的人胚神经干细胞(human neural stem cells,hNSCs)对大鼠损伤脊髓治疗机制和治疗的作用机制。方法在体外用Lenti- virus构建同时表达绿色荧光蛋白(green fluorescence protein,GFP)和NT-3的hNSCs,将其移植到T_(10)脊髓半横断损伤的大鼠体内,用荧光显微镜和BBB评分观察体内转基因表达和大鼠功能恢复情况,用免疫组化染色检测移植细胞在体内存活分化情况。结果(1)在体外用Lentivirus成功构建了同时表达多基因的人胚基因工程NSC并检测到了稳定的转基因表达;(2)用荧光显微镜可以检测到到hNSC,能在体内长时间存活并表达转基因;(3)BBB评分检测到hNSCs移植组大鼠后肢功能有明显恢复;(4)移植的基因工程hNSC能在体内分化为神经元及星形胶质细胞等功能细胞。结论Lentivirus介导NT-3转染的hNSC能够调整损伤的局部微环境促进损伤脊髓的功能恢复。

Study of genetic engineering human neural stem cells modified by lentivirus to deliver NT-3 for repair of spinal cord injury in rats

Objective To explore the therapeutic mechanism and effect of genetic engineering human neural stem cells (NSC) modified by Lentivirus to deliver neurotrophin 3 (NT-3) for the treatment of spinal cord injury in rat.Methods Genetic engineering NSC modified by Lentivirus to deliver both GFP ( green fluorescence protein) and NT-3 in vitro was constructed and identified.The genetic engineer- ing NSC was transplanted into the rat spinal cord after T_(10) hemisection,then the transgenic expression in vivo was detected using fluorescence microscope,the function recovery was estimated using BBB,and the survival and differentiation of the transplanted cells in vivo were detected with immunohistochemistry. Results The genetic engineering human NSC modified by Lentivirus to deliver NT-3 was successfully constructed.There was a steady expression of objective genes in vitro;the genetic engineering human NSC could survive and express the transferred genes stably for a long time in vivo by using fluorescence microscope.The hind-limb function of the injured rat acquired obvious improvement,and the transplanted hNSC could survive for a long time and differentiate into neurons and astrocytes in vivo;Conclusion The genetic engineering NSC modified by Lentivirus to deliver NT-3 can promote the functional recovery of injured spinal cord by means of improving the local microenvironment of injured spinal cord.